Interleukin-17A is a newly described proinflammatory cytokine, which plays important roles in autoimmune diseases as well as asthma. In current work, we constructed a recombinant plasmid pMFA42S-Ag85a-IL-17a by inserting fusion gene Ag85a-IL-17a into shuttle vector pMFA42S, which was transformed to Mycobacterium smegmatis by electroporation to obtain recombinant M. smegmatis named rMS-Ag85a-IL-17a. The comparison of growth pattern between M. smegmatis and rMS-Ag85a-IL-17a suggested fusion gene had no significant influence on the growth of strains, and rMS-Ag85a-IL-17a expressed fusion protein Ag85A-IL-17A which had good immunogenicity revealed by Western blot. M. smegmatis and rMS-Ag85a-IL-17a were performed to intranasally immunize mice; then, antibody response in sera was detected by enzyme-linked immunosorbent assay. Our findings demonstrated that rMS-Ag85a-IL-17a could induce specific IL-17A autoantibody in mice, which laid the foundation for further study.